Isolation and characterization of cDNA clones for the gamma subunit of Xenopus fibrinogen, the product of a coordinately regulated gene family.
Fibrinogen, the major structural protein involved in blood coagulation, is synthesized and secreted by the liver. In the frog Xenopus laevis, fibrinogen production is dramatically induced by glucocorticoids. The hormonal stimulation requires synthesis of three separate subunits, designated A alpha, B beta, and gamma. For investigation of the molecular mechanisms underlying this coordinate induction, we have isolated cDNA clones for the subunits of Xenopus fibrinogen. In this communication we describe the identification of clones for the gamma chain. Initially, a Xenopus liver cDNA library in pBR322 was screened with a rat gamma chain cDNA and a clone representing half of the 1600-base frog gamma mRNA was identified. This clone was shown to be complementary to gamma mRNA by hybrid selection of mRNA that translated in vitro into the gamma polypeptide. A clone about 1460 base pairs in length was then isolated from a Xenopus liver lambda gt10 cDNA library and subcloned into Bluescript SK-. This clone, designated X1 gamma 3, contains the entire 3'-end and lacks 38 bases at the 5'-end of gamma mRNA. The deduced amino acid sequence at the N-terminal is compatible with a signal peptide of 20-23 amino acids, in agreement with the calculated size of the frog gamma chain signal peptide. Following the signal sequence is a region of highly conserved amino acids that participate in disulfide bond formation critical for the maintenance of tertiary structure in mammalian fibrinogen. The gamma cDNA clone was used to measure gamma mRNA in purified Xenopus liver cells treated with glucocorticoids in primary culture.(ABSTRACT TRUNCATED AT 250 WORDS)
Bhattacharya
A
A
Department of Physiology and Biophysics, University of Iowa College of Medicine, Iowa City 52242.
Shepard
A R
AR
Moser
D R
DR
Holland
L J
LJ
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D004247:DNA / Q000235:genetics*; D003907:Dexamethasone / Q000494:pharmacology; D005340:Fibrinogen / Q000235:genetics*; D005786:Gene Expression Regulation / Q000187:drug effects; D005796:Genes; D006801:Humans; D007798:Lampreys / Q000235:genetics; D008099:Liver / Q000737:chemistry; D008969:Molecular Sequence Data; D005810:Multigene Family; D021382:Protein Sorting Signals / Q000235:genetics; D051381:Rats; D012689:Sequence Homology, Nucleic Acid; D014982:Xenopus laevis / Q000235:genetics*
D021382:Protein Sorting Signals; D003907:Dexamethasone; D005340:Fibrinogen; D004247:DNA